Effect Of Perampanel Monotherapy On Pentylenetetrazol-kindling In Mice And Its Influence On Cognitive Function

Objective:The epileptic model of mice kindled by pentylenetetrazol(PTZ)was established to explore the effects of low-dose,medium-dose and high-dose perampanel(PER)on seizure grade,serum glutamic acid content,glial fibrillary acidic protein(GFAP)-positive astrocytes and electroencephalogram(EEG)in PTZ-kindled mice,as well as the differences of escape latency and spatial exploration ability in Morris water maze experiment so as to evaluate the anti-epileptic effect of different doses of PER and analyze the influence of PER on cognitive impairment in anti-epileptic treatment.Methods:Fifty clean and healthy Balb/c male mice(5-9 weeks old,equivalent to 4-7 years old in humans)were randomly divided into five groups,namely,control group(CON group),pentylenetetrazol group(PTZ group),low-dose PER group(L-PER group),middle-dose PER group(M-PER group)and high-dose PER group(H-PER group),with 10 mice in each group.At first,the L-PER group,M-PER group and H-PER group were given 0.75,1.5 and 3 mg/kg/d PER by gavage(concentrations: 0.1 mg/ml,0.2 mg/ml and 0.4 mg/ml,volume: 7.5 ml/kg/d)while the PTZ group and the CON group were given the same volume of normal saline(7.5 ml/kg/d)by gavage.After 30 minutes,the PTZ group,LPER group,M-PER group and H-PER group were given 35 mg/kg/d PTZ(concentration:2mg/ml,volume: 17.5 ml/kg/d)by intraperitoneal injection while the CON group was given the same volume of normal saline(17.5 ml/kg/d)by intraperitoneal injection,and then we observed the behavioral performance of all mice for 60 minutes.Once a day for4 weeks.On the 28 th day of treatment,the latency,grade and duration of epileptic seizures of mice in each group were observed,and the scores were recorded according to Racine grading standard,and water maze experiment(positioning navigation experiment and space exploration experiment)was completed to detect the changes of cognitive function,and EEG was recorded.Then,serum and brain samples of mice in each group were taken to detect the content of serum glutamic acid by mouse ELISA glutamic acid kit and the area ratio of glial fibrillary acidic protein(GFAP)positive astrocytes in brain by immunohistochemical method.Results:1.As for the behavioral performance,the L-PER group,M-PER group and H-PER group have significantly prolonged the latency of epileptic seizures,reduced the seizure grade and duration compared with the PTZ group(P < 0.05).The anti-epileptic effect of the M-PER group and H-PER group were better than that of the L-PER group(P < 0.05).The anti-epileptic effect between the M-PER group and H-PER group were equivalent(P > 0.05).2.As for the cognitive function,the escape latency increased and the times of crossing the original platform position reduced and the time of staying in the target quadrant increased significantly of mice in the PTZ group compared with the CON group(P < 0.05).The escape latency and the time of staying in the target quadrant decreased and the times of crossing the original platform position increased significantly of mice in the L-PER group,M-PER group and H-PER group compared with the PTZ group(P < 0.05).Compared with the L-PER group,M-PER group and H-PER group reduced the escape latency and the time of staying in the target quadrant and increased the number of times of crossing the original platform position significantly(P < 0.05).There was no significant difference in cognitive function between the M-PER group and H-PER group(P > 0.05).3.As for the EEG,the EEG activity of the L-PER group,M-PER group and H-PER group were improved during the seizure compared with the PTZ group.4.As for the serum glutamic acid,compared with the CON group,the content of serum glutamic acid in the PTZ group increased significantly(P < 0.01).That in the LPER,M-PER and H-PER group decreased significantly compared with the PTZ group(P< 0.01).The content of serum glutamic acid in the M-PER and H-PER group were lower than that in the L-PER group(P < 0.01).There was no significant difference between the M-PER and H-PER group(P > 0.05).5.As for the expression of astrocytes,the area ratio of astrocytes in the PTZ group increased significantly compared with the CON group(P < 0.01).Compared with the PTZ group,The area of astrocytes in the L-PER group,M-PER group and H-PER group decreased significantly(P < 0.01).The area of astrocytes in the M-PER group and H-PER group were lower significantly than that in the L-PER group(P < 0.01).There was no significant difference between the M-PER group and H-PER group(P > 0.05).Conclusion:1.PER has antiepileptic effect on mice kindled by PTZ and PER with a dose of1.5mg/kg/d can show the greatest antiepileptic effect.2.In the antiepileptic treatment of PTZ-kindled mice,PER has no damage to cognitive function and can even improve cognitive function and increasing the dose of PER can not aggravate the cognitive dysfunction.